Figure 5
From: Highly purified mussel adhesive protein to secure biosafety for in vivo applications
Assessment of biological safety of purified recombinant MAP using inflammatory response assays on RAW 264.7 macrophage cells. (A) TNF-α and (B) IL-6 mRNA expressions were observed by treating MAP samples at each purification step. The data represent mean +/− standard deviation and were analyzed 2-tails student t-test (*p < 0.05, **p < 0.01, ***p < 0.005). Abbreviations: NC, non-treated negative control; WC, whole cells; IB, isolated inclusion body containing MAP after disruption of CaCl2/EDTA-treated whole cells; X-100/100, acetic acid-extracted MAP after inclusion body washing with Triton X-100 twice; X-100/114, acetic acid-extracted MAP after inclusion body washing with Triton X-100 and Triton X-114 in order; X-114/100, acetic acid-extracted MAP after inclusion body washing with Triton X-114 and Triton X-100 in order; X-114/114, acetic acid-extracted MAP after inclusion body washing with Triton X-114 twice; IE: MAP fraction through cation exchange chromatography of acetic acid-extracted solution after inclusion body washing with Triton X-114 twice; ppMAP, purified MAP based on previous method; LPS 1EU, LPS standard with 1 endotoxin unit; LPS 5EU, LPS standard with 5 endotoxin unit; LPS 10EU, LPS standard with 10 endotoxin unit.